KR20100041175A - N1-2-tiopen-2-ylethyl-n2-substituted biquanide derivatives, method for the preparation thereof and pharmaceutical composition comprising the same - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing N1-2-thiopene-2-ylethyl-N2-substituted biguanide derivative is provided to ensure function of reducing blood sugar and lipid and to prevent and treat diabetes, metabolic syndrome, or p53-deficient cancer. CONSTITUTION: A N1-2-thiopene-2-ylethyl-N2-substituted biguanide derivative compound is denoted by chemical formula 1. In chemical formula 1, R is C1-C8 alkyl, aryl, C1-C8 alkoxyalkyl, C3-C7 cycloalkyl, C3-C7 cycloalkylC1-C3 alkyl, substituted phenyl or phenyl C1-C3 alkyl, substituted naphthyl or naphthyl C1-C3 alkyl. A method for preparing a compound of chemical formula 1 comprises: a step of reacting a compound of chemical formula 2 with NaBH4 to obtain a compound of chemical formula 3; a step of reacting the compound of chemical formula 3 with thionylchloride in organic solvent to obtain a compound of chemical formula 4; a step of reacting the compound of chemical formula 4 with sodium cyanide in DMSO to obtain a compound of chemical formula 5; a step of reacting the compound of chemical formula 5 with sodium borohydride and nickel chloride hydrate to obtain a compound of chemical formula 6; a step of reacting the compound of chemical formula 6 with R-isothiocyanate(RNACS) under the presence of base in organic solvent to obtain a compound of chemical formula 8; and a step of reacting the compound of chemical formula 8 in guanidine solution under the presence of mercury oxide.

Description

N1-2-thiophen-2-ylethyl-N2-substituted biguanide derivatives, a preparation method thereof, and a pharmaceutical composition containing the same as an active ingredient {N1-2-TIOPEN-2-YLETHYL-N2-SUBSTITUTED BIQUANIDE DERIVATIVES, METHOD FOR THE PREPARATION THEREOF AND PHARMACEUTICAL COMPOSITION COMPRISING THE SAME}

The present invention provides a N1-2-thiophen-2-ylethyl-N2-substituted biguanide derivative, which shows excellent hypoglycemic activity and lipid lowering activity even at a lower dose than a conventional drug, a method for preparing the same, and an active ingredient It relates to a pharmaceutical composition containing.

Diabetes mellitus is a disease characterized by persistent hyperglycemic symptoms. Carbohydrate and lipid metabolic disorders are the main conditions, and systemic complications due to hyperglycemia and decreased glucose utilization are worsening. This diabetic condition is caused by insulin deficiency or insulin resistance, and diabetes caused by insulin resistance is called type 2 diabetes.

Type 2 diabetes is caused by a condition in which insulin is unable to carry sugar into the cell due to a decrease in insulin receptors or a deficiency in the signaling system through the receptor, that is, a condition in which insulin develops resistance, It exacerbates direct vascular destruction and metabolic syndrome due to hyperinsulinemia.

Many types of diabetes drugs have been used to treat this type 2 diabetes. However, drugs other than biguanide metformin have some effects on hypoglycemic effects, but they are not effective in preventing serious complications such as vision loss, heart failure, stroke, kidney failure, peripheral nerve disorders, and foot ulcers. It was. For example, sulfonyurea drugs can be found in the pancreas. It is a drug that lowers blood sugar by forcibly secreting insulin, and the effect of the drug disappears quickly, and abnormal lipid metabolism causes arteriosclerosis, weight gain, and brain damage due to hypoglycemia. In addition, the glitazone-based drug mainly needs to be administered in combination with metformin to solve only the problem of insulin resistance to adipose tissue, causing side effects such as retinal vessel destruction, so there is a disadvantage that requires very careful use.

Metformin, on the other hand, has the same effect as insulin, and does not cause hypoglycemia, solves the problem of insulin resistance in adipose tissue, liver tissue and muscle tissue. It exhibits a glycosylated hemoglobin level lowering effect. It is also reported to activate AMP-activated protein kinase (AMPK) to normalize hyperglycemia, improve lipid status, normalize menstrual irregularities, ovulation and pregnancy, treat fatty liver and even prevent cancer.

However, metformin is generally administered three times a day, and since a single dose is about 500 mg or more, a tablet containing about 1,500 mg or more of metformin is required to sustained release it for a single daily use, in which case the size of the tablet is too large. I have difficulty taking it. In addition, the current long-acting formulations contain only about 750 mg metformin in one tablet, so there is a problem in that two or more tablets should be taken when administering. Therefore, there is an urgent need for the development of a formulation that can reduce the dose of drugs required to exert an excellent hypoglycemic and hypolipidemic effect.

It is an object of the present invention to provide a novel compound and a method for preparing the same, which exhibit excellent blood glucose lowering action and lipid lowering action even at a lower dose than conventional drugs.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating metabolic syndrome such as diabetes mellitus, insulin-independent diabetes mellitus, obesity and atherosclerosis, or a cancer lacking the gene P53.

In order to achieve the above object, the present invention provides a N1-2-thiophen-2-ylethyl-N2-substituted biguanide derivative compound of Formula 1, or a pharmaceutically acceptable salt thereof:

Where

R is C ₁ -C ₈ alkyl, allyl, C ₁ -C ₈ alkoxyalkyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl (C ₁ -C ₃ ) alkyl, optionally substituted phenyl or phenylC _1- C ₃ alkyl, optionally substituted naphthyl or naphthylC ₁ -C ₃ alkyl, or adamantyl group;

R ₁ is hydrogen or a C ₁ -C ₆ alkyl group.

In addition, the present invention provides a method for preparing the compound of Formula 1.

The present invention also provides a pharmaceutical composition for preventing or treating diabetes, comprising the compound of Formula 1 as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating metabolic syndrome comprising the compound of Formula 1 as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer lacking the gene P53 comprising the compound of Formula 1 as an active ingredient.

The N1-2-thiophen-2-ylethyl-N2-substituted biguanide derivative compound of Formula 1 exhibits hypoglycemic action and hypolipidemic activity, and excellent hypoglycemic action and lipid lowering even at low doses It can be used for the prevention or treatment of metabolic syndrome such as diabetes mellitus, insulin independent diabetes mellitus, obesity and atherosclerosis, and cancer lacking gene P53.

Hereinafter, the present invention will be described in detail.

In the compound of Formula 1 according to the present invention, "C ₁ -C ₈ alkyl" includes a linear or branched alkyl group, preferred examples thereof are methyl, ethyl, n-propyl, isopropyl, n-butyl, iso Butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl and n-octyl.

As used herein, "C ₁ -C ₈ alkoxyalkyl" includes methoxymethyl, methoxyethyl, methoxyethoxyethoxyethyl, ethoxymethyl, octyloxymethyl and the like.

Preferred examples of “C ₃ -C ₇ cycloalkyl” as used herein include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

Preferred examples of "C ₃ -C ₇ cycloalkylC ₁ -C ₃ alkyl" as used herein include cyclopropanemethyl, cyclobutanemethyl, cyclopentanemethyl, cyclohexanemethyl, cyclopropaneethyl, cyclobutaneethyl, cyclopentaneethyl And cyclohexane ethyl, cyclopropanepropyl, cyclobutanepropyl, cyclopentanepropyl, and cyclohexanepropyl.

Preferred examples of “optionally substituted phenyl or phenylC ₁ -C ₃ alkyl” include benzyl, phenyl, naphthyl, 1-phenylethyl, 2-phenylethyl, 1-phenylpropyl unsubstituted or substituted with a substituent. , 2-phenylpropyl and 3-phenylpropyl. The substituents may be the same or different from 1 to 6, preferably 1 to 3, respectively, and these substituents may be substituted at any position chemically acceptable to phenyl. Examples of the substituent include a halogen atom, hydroxy group, nitro group, cyano group, C ₁ -C ₆ alkyl group, C ₁ -C ₆ haloalkyl group, C ₁ -C ₆ cycloalkyl group, C ₆ -C ₁₀ aryl group, C _6- C ₁₀ aryloxy group, C ₁ -C ₆ alkoxy group, C ₁ -C ₆ haloalkoxy group, C ₃ -C ₆ cycloalkyloxy group, C ₁ -C ₇ alkanoyl group, carboxyl group, carbamoyl group, alkylamino group , C ₂ -C ₇ sulfonic acid group, sulfonamido group and C ₁ -C ₆ alkylthio group.

Preferred examples of “optionally substituted naphthyl or naphthylC ₁ -C ₃ alkyl” as used herein include 1-naphthylmethyl, 2-naphthylmethyl, 1-naphthylethyl and 2, unsubstituted or substituted with substituents. - may be made of naphthyl acetate, is the same as the substituent for the "phenyl or phenyl-C ₁ -C ₃ alkyl" as a substituent.

Preferred examples of “C ₁ -C ₆ alkyl” as used herein include methyl, ethyl propyl, butyl, pentyl and hexyl.

In Formula 1, more preferably R is ethyl, t-octyl, 1-naphthyl, hexyl, methyl, cyclohexyl, t-butyl, benzyl, 2-ethylphenyl, propyl, 3-trifluoromethylphenyl , 4-methoxyphenyl, 2,5-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 4-fluorophenyl, 4-methylphenyl, phenyl, cyclopentyl, cycloheptyl, pentyl, methoxy Ethyl, butyl, 3-methylphenyl, 4-t-butylphenyl, 3-hexylphenyl, 4-methoxyphenyl, 2,5-dimethoxyphenyl, 3,5-dimethoxyphenyl, 4-bromophenyl, 3-bromophenyl, 3-phenylphenyl, 3-phenoxyphenyl, 4-bromobenzyl, 4-methoxybenzyl, 3,5-dichlorophenyl, 4-methylbenzyl, isopropyl, isobutyl, cyclopropyl Methyl, or allyl; R ₁ is hydrogen, 3-methyl or 5-ethyl.

Pharmaceutically acceptable salts of the compounds of formula 1 according to the invention are organic acids, such as formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, Succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid salts; Inorganic acids such as hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid salts. The above-mentioned acid addition salts can be, for example, a) directly mixing the compound of formula 1 and acid, b) dissolving one of them in a solvent or a hydrous solvent, or c) compound of formula 1 and The acid can be prepared by placing it in an acid in a solvent or a hydration solvent and applying it to a general method for preparing a salt thereof.

When the compound of Formula 1 has an acidic group such as a carboxyl group and a sulfonic acid group, the compound is a zwitterionic salt, and such salts are alkali metal salts (such as sodium salts and potassium salts), alkaline earth metal salts (such as calcium salts and Magnesium salts, etc.), inorganic acid salts such as aluminum salts and ammonium salts, and basic addition salts such as trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclo Hexylamine and N, N'-dibenzylethylenediamine salt). In addition, the salt of the compound of Formula 1 may be a basic amino acid salt (eg arginine, lysine and ornithine salt) and acidic amino acid salt (eg aspartic acid salt). The salt of the compound of formula 1 is preferably a pharmaceutically acceptable salt, more preferably an acid addition salt, even more preferably acetate, hydrochloride, hydrobromide, methanesulfonate, malonate or oxalate.

For example, the method for preparing the compound of Formula 1 according to the present invention,

1) reacting a compound of Formula 2 with NaBH ₄ in a solvent to obtain a compound of Formula 3;

2) reacting a compound of Formula 3 with thionyl chloride in an organic solvent to obtain a compound of Formula 4;

3) reacting the compound of Formula 4 with sodium cyanide in DMSO to obtain a compound of Formula 5;

4) reacting the compound of Formula 5 with sodium borohydride and nickel chloride hydrate in a solvent to obtain a compound of Formula 6;

5) reacting a compound of Formula 6 with R-isothiocyanate (RNCS) in an organic solvent in the presence of a base to obtain a compound of Formula 8; And

6) reacting a compound of formula 8 in a guanidine solution in the presence of mercury oxide:

[Formula 1]

In the above formula, R and R ₁ are as defined in the formula (1).

As another method, the compound of formula 1 of the present invention

1) reacting a compound of Formula 2 with NaBH ₄ in a solvent to obtain a compound of Formula 3;

2) reacting a compound of Formula 3 with thionyl chloride in an organic solvent to obtain a compound of Formula 4;

3) reacting the compound of Formula 4 with sodium cyanide in DMSO to obtain a compound of Formula 5;

4) reacting the compound of Formula 5 with sodium borohydride and nickel chloride hydrate in a solvent to obtain a compound of Formula 6;

5) reacting a compound of formula 6 with ethylchloroformate in an organic solvent in the presence of carbon disulfide and a base to obtain a compound of formula 7;

6) reacting a compound of Formula 7 with NH ₂ R in a solvent in the presence of a base to obtain a compound of Formula 8; And

7) reacting the compound of formula 8 in a guanidine solution in the presence of mercury oxide:

[Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Chemical Formula 5]

[Formula 6]

[Formula 8]

In the above formula, R and R ₁ are as defined in the formula (1).

The preparation methods may be represented by, for example, the following Scheme 1, which is described step by step as follows:

In the above formula, R and R ₁ are as defined in the formula (1).

In step (1), sodium borohydride (NaBH ₄ ) and a solvent (e.g., methanol, ethanol, propanol, isopropanol, butanol, tetra) are used as starting materials of the carboxaldehyde compound of substituted thiophene of formula (2). Hydrofuran, acetonitrile, N, N-dimethylformamide or dimethylsulfoxide) may be reacted at 0 ° C. to reflux temperature to obtain an alcohol derivative of formula 3. The amount of sodium borohydride used at this time is 1 to 3 molar equivalents relative to the compound of formula (2).

 In this case, the compound of Formula 2 used as a starting material in the reaction can be easily synthesized by using a commercially available compound, or by a known method.

In step (2), the compound of formula 3 obtained above is reacted with thionyl chloride in an organic solvent (for example, dichloromethane, dichloroethane or dimethylformamide) at room temperature to reflux temperature range of Chloride derivatives can be obtained. The amount of thionyl chloride used is 3 to 10 molar equivalents relative to the compound of formula (3).

In step (3), the compound of formula 4 obtained above is reacted at room temperature in sodium cyanide and dimethyl sulfoxide (DMSO) to obtain a cyanide derivative of formula (5). The amount of sodium cyanide used at this time is 2 to 4 molar equivalents relative to the compound of formula (4).

In step (4), the compound of formula 5 obtained above is added with sodium borohydride, nickel chloride hydrate and a solvent (e.g., methanol, ethanol, propanol, isopropanol, butanol, tetrahydrofuran, acetonitrile, N, N -Dimethylformamide or dimethylsulfoxide) at room temperature to give a substituted thiophene ethylamine derivative of formula (6). The amount of sodium borohydride used at this time is 3 to 5 molar equivalents relative to compound 5, and the amount of nickel chloride hydrate is 1 to 2 molar equivalents relative to compound 5.

Subsequently, the thiourea compound of formula (8), which is used as an intermediate in the preparation method of the compound of formula (1), may be substituted with thiophene ethylamine of formula (6) when isothiocyanate (RNCS) having a substituent R group is commercially available. When reacted with RNCS in a solvent in the presence of a base, or (B) RNCS bearing a substituent R group is not commercially available, thiophene ethylamine of formula 6 is substituted with ethylchloro in an organic solvent in the presence of carbon disulfide and a base. After reacting with formate to convert the isothiocyanate compound of formula 7, the compound of formula 7 can be obtained by reacting with NH ₂ R in an organic solvent in the presence of a base.

As the base used in the steps (5), (5 ′) and (6) when preparing the thiophene ethylamine compound of Chemical Formula 6, triethylamine, trimethylamine or diisopropylethylamine may be used, and the organic solvent may be used. As the dichloromethane, dichloroethane or dimethylformamide may be used. The reaction temperature is in the range from 0 ° C to room temperature. The amounts of carbon disulfide, base and ethylchloroformate in the above steps (5) and (5 ') can be used in about 1 to 2 molar equivalents relative to the compound of formula (6), respectively.

In step (7), the thiourea compound of formula (8) obtained above is dissolved in mercury oxide and a suitable organic solvent (for example, ethyl alcohol, methyl alcohol or dimethyl formamide), and then refluxed by adding 1M guanidine ethanol solution. In this case, the amount of mercury oxide is about 1 to 2 molar equivalents relative to compound 8, the 1M guanidine ethanol solution is 1 to 3 molar equivalents relative to compound 8, and the reaction temperature is in the range up to the reflux temperature of the solvent used (for example, dimethyl). In the case of formamide). After completion of the reaction, the resultant is filtered and, for example, by adjusting the pH of the reaction solution to about 4 to 5 using an acid such as hydrochloric acid, and concentrating and purifying the resulting solution. Alternatively, acceptable salts can be obtained.

The compound of Formula 1 thus obtained or a pharmaceutically acceptable salt thereof exhibits hypoglycemic action and hypolipidemic action even at a lower dose than conventional drugs. It can be usefully used for the prevention and treatment of metabolic syndrome.

Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of metabolic syndrome comprising the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. Examples of the metabolic syndrome include insulin-independent diabetes, obesity, atherosclerosis, and the like.

In another aspect, the present invention provides a pharmaceutical composition for preventing or treating diabetes mellitus as an active ingredient of the compound of formula 1 or a pharmaceutically acceptable salt thereof.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer, which lacks the gene P53 having the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound of formula 1 or a salt thereof may be administered on its own or in a pharmaceutical composition comprising one or more additives. The additives include, for example, stabilizers, binders, bases, dragees, excipients, disintegrants, dissolution aids, viscosity regulators, coating agents, suspending agents, pH adjusting agents, isotonic agents, brightening agents, emulsifiers, sweeteners, copulating agents, wetting agents, Choose from soft capsule base, hard capsule base, plasticizer, preservative, antioxidant, solvent, lubricant, pressure-sensitive adhesive, surfactant, light-shielding agent, colorant, dispersing agent, analgesic agent, buffer, refreshing agent, and solubilizer according to administration route and compounding purpose. Or used in combination.

The pharmaceutical compositions can be prepared using formulation techniques that can be practiced by those skilled in the art to suit their particular use and desired purpose.

The pharmaceutical compositions of the invention can be administered orally, e.g. in the form of tablets, powders, granules, pills, troches, capsules, liquids. It may also be administered parenterally, for example, in the form of injections, suppositories, ointments, external preparations, pastes, cataplasmas, linings, patches.

Suitable dosage levels for oral administration of the compound of formula 1 or a pharmaceutically acceptable salt thereof according to the invention are preferably in the range of 0.5 to 150 mg per kg based on 60 kg adult males once daily. However, this may vary depending on various factors such as the age and symptoms of the patient, the route of administration, and in some cases, may be administered in an amount less than or greater than the dosage range.

Hereinafter, the present invention will be described in more detail with reference to Examples, which are only intended to assist in understanding the configuration and operation of the present invention, and the scope of the present invention is not limited to these Examples.

Example

Example 1: Preparation of N1- (thiophen-2-ylethyl) -N2-ethyl biguanide hydrochloride

(1-1) Synthesis of 1-ethyl-3- (2- (thiophen-2yl) ethyl) thiourea

Ethyl isothiocyanate (1.3 g, 15.0 mmol) was slowly added dropwise to a thiophenethylamine (1.3 g, 10.2 mmol) -containing dichloromethane (150 mL) solution. Triethylamine (3.8 mL, 20.4 mmol) was added dropwise to the mixture, followed by stirring at room temperature for 2 hours. After the reaction was completed, the pH of the reaction solution was adjusted to about 7 with 1N HCl, and water was added to extract dimethyl chloride. Flash column chromatography (ethylacetate: hexane = 1: 3) recovered 1-ethyl-3- (2- (thiophen-2yl) ethyl) thiourea (1.9 g, 89%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.45 (br s, 1 H), 7.42 (br s, 1 H), 7.39-7.30 (m, 1 H), 6.95-6.97 (m, 1 H), 6.87-6.88 (m, 1H), 3.60-3.50 (m, 2H), 3.40-3.25 (m, 2H), 3.00 (t, 2H, J = 7.4), 1.02 (t, 3H, J = 7.2)

(1-2) Synthesis of N1- (thiophen-2-ylethyl) -N2-ethyl biguanide hydrochloride

Mercury oxide (Mercuric oxide (II), 2.2 g, 10.2 mmol) was prepared in step 1-1, 1-ethyl-3- (2- (thiophen-2yl) ethyl) thiourea (1.1 g, 5.10 mmol). -To ethanol (15 mL) solution was added. To the reaction solution was slowly added dropwise 1M guanidine ethanol solution (15 mL, 15.3 mmol), followed by reflux for 12-24 hours. After the reaction solution was cooled, the reaction solution was filtered using a celite filter, and 2N HCl was added to the filtrate, and the mixture was titrated to pH 4-5. After concentration of the solution, flash column chromatography (dichloromethane: methanol = 9: 1) was used to recover N1- (thiophen-2-ylethyl) -N2-ethyl biguanide hydrochloride (0.21 g, 15 %).

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.33-7.34 (m, 1H), 6.85-6.94 (m, 2H), 3.28-3.32 (m, 2H), 2.96-3.11 (m, 4H), 0.99- 1.04 (m, 3 H); mp 128.0-131.7 ℃

Examples 3 to 5, by the same method as in Example 1, except for using the isothiocyanate compound corresponding to the target compound instead of ethyl isothiocyanate in step (1-1) of Example 1 7, 8, 10 to 13, 20, 23, 25, 26, 41 and 42 were prepared .

Example 2: Preparation of N1-2- (thiophen-2-ylethyl) -N2-t-octyl biguanide hydrochloride

(2-1) Synthesis of 2- (2-isothiocyanatoethyl) thiophene

The thiophene-2-ethylamine (4.60 mL, 39.31 mmol) and triethylamine (5.00 mL, 39.31 mmol) -containing 1,2-dichloroethane (25 mL) solutions were cooled to 0 ° C. and then 1,2 -A solution of carbon disulfide (2.36 mL, 39.31 mmol) in dichloroethane (50 mL) was added dropwise over 15 minutes. The reaction solution was raised to room temperature and then cooled to 0 ° C., and ethylchloroformate (4.27 mL, 31.39 mmol) was added dropwise. Then, after stirring for 1-2 hours at room temperature, water (150 mL) and 2N sodium hydroxide solution (75 mL) were added, followed by extraction with dichloromethane. The combined extracts were washed with water and brine, dried over sodium sulfate and concentrated to recover 2- (2-isothiocyanatoethyl) thiophene as a yellow liquid (5.37 g, yield: 80.75%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.22 (dd, 1H, J = 5.1, 1.2 Hz, Thiophene), 6.98 (dd, 1H, J = 5.1, 3.4 Hz, Thiophene), 6.93-6.91 (m, 1H , Thiophene), 3.76 (t, 2H, J = 6.7 Hz, CH 2), 3.22 (t, 2H, J = 6.8 Hz, CH 2)

(2-2) Synthesis of 1-t-octyl-3- (2- (thiophen-2yl) ethyl) thiourea

2 (2-Isothiocyanatoethyl) thiophene (2.5 g, 15.0 mmol) and t-octylamine (1.6 mL, 10.2 mmol) were used in the same manner as in Step 1-1, except that 1 -t-octyl-3- (2- (thiophen-2yl) ethyl) thiourea was synthesized (2.5 g, 83%).

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.33 (m, 1H), 7.31 (br s, 1H), 7.19 (br s, 1H), 6.93 (m, 1H), 6.84 (m, 1H), 3.57 (m, 2H), 2.96 (t, 2H, J = 7.0), 1.97 (s, 2H), 1.97 (s, 6H), 0.92 (s, 9H),

(2-3) Synthesis of N1-2- (thiophen-2-ylethyl) -N2-t-octyl biguanide hydrochloride

The same method as in Step 1-2, except that 1-t-octyl-3- (2- (thiophen-2yl) ethyl) thiourea (1.5 g, 5.10 mmol) prepared in Step 2-2 was used. To recover N1-2- (3-methylthiophen-2-ylethyl) -N2-t-octyl biguanide hydrochloride (0.31 g, 17%).

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.33-7.36 (m, 1H), 6.89-6.96 (m, 2H), 3.20-3.21 (m, 2H), 2.97-2.99 (m, 2H), 1.28- 1.30 (m, 6H), 0.90-0.93 (m, 11H)

Example 6, 9, 14 to 19, by the same method as Example 2 except for using the amine compound corresponding to the target compound instead of tert-octylamine in the step (2-2) of Example 2 The compounds of 21, 22, 24, 27 to 40, 43 and 44 were prepared.

Example 45 Preparation of N1-2- (3-methylthiophen-2-ylethyl) -N2-methyl biguanide hydrochloride

(45-1) Synthesis of (3-methylthiophen-2-yl) methanol (Compound 3)

The 3-methyl-2-thiophene-carbosaaldehyde (1.00 g, 7.93 mmol) -containing methanol (16 ml) solution was cooled to 0 ° C. and then sodium borohydride (0.60 g, 15.86 mmol) was added. After stirring the reaction solution for 30 minutes at room temperature, water (30 ml) was added and extracted with ethyl acetate. The organic layers were collected, washed with brine, dried over sodium sulfate and concentrated to give column chromatography (ethyl acetate: hexane = 20: 80). ) Was obtained (3-methylthiophen-2-yl) methanol (0.89 g, 88%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.17 (d, 1H, thiophene), 6.84 (d, 1H, thiophene), 4.76 (d, 2H, J = 4.92 Hz, CH 2), 2.25 (s, 3H, CH 3 ), 1.66 (t, 1H, J = 5.43 Hz, OH); IR (neat): 3320, 3100, 3060, 2918, 2861, 1555, 1430, 1383, 1368, 1353, 1300, 1231, 1171, 1032, 996, 932, 876, 832, 703, 663 cm-1; MS (ESI +) m / z 128.19 [M < + >].

(45-2) Synthesis of 2- (3-methylthiophen-2-yl) acetonitrile (Compound 5)

Thionyl chloride (0.85 ml, 11.70 mmol) was slowly added to the (3-methylthiophen-2-yl) methanol (0.50 g, 3.90 mmol) -containing dichloromethane (5 ml) solution prepared in step 45-1. It was added dropwise, stirred at room temperature for 30 minutes, and then concentrated under reduced pressure to give chloride (Compound 4). The concentrated solution obtained above was dissolved in dimethyl sulfoxide (2 ml), sodium cyanide (0.57 g, 11.70 mmol) was added thereto, and the mixture was stirred at room temperature for 1 hour. Water (30 ml) was added to the reaction solution, which was then extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, concentrated and separated by column chromatography (ethyl acetate: hexane = 20: 80) to give 2- (3-methylthiophen-2-yl) acetonitrile (0.39 g, 56%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.16 (d, 1H, thiophene), 6.84 (d, 1H, thiophene), 3.78 (s, 2H, CH ₂ ), 2.24 (s, 3H, CH ₃ ); IR (neat): 3098, 2917, 2248, 1557, 1432, 1409, 1383, 1367, 1298, 1237, 1159, 1083, 1023, 952, 915, 879, 855, 828, 738, 706, 664 cm ^-1 .

(45-3) Synthesis of 2- (3-methylthiophen-2-yl) ethanamine (Compound 6)

Nickel chloride (0.95 g, 0.73 mmol) was added to the 2- (3-methylthiophen-2-yl) acetonitrile (0.10 g, 0.73 mmol) -containing ethanol (3 mL) solution prepared in step 45-2. It was. Sodium borohydride (0.83 g, 2.19 mmol) was added in small portions to the reaction solution and stirred at room temperature. Water (30 ml) was added to the reaction solution, which was then extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, concentrated and separated by column chromatography (dichloromethane: methanol = 8: 2) to obtain 2- (3-methylthiophen-2-yl) ethanamine ( 51 mg, 50%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.09 (d, 1H, thiophene), 6.80 (d, 1H, thiophene), 3.21 (s, 4H, 2 × CH 2), 2.21 (s, 3H, CH 3).

(45-4) Synthesis of N1-2- (3-methylthiophen-2-ylethyl) -N2-methyl thiourea

The above step except that methyl isothiocyanate (1.0 mL, 15.0 mmol) and 2- (3-methylthiophen-2-yl) ethanamine (1.4 g, 10.2 mmol) prepared in step 45-3 were used N1-2- (3-methylthiophen-2-ylethyl) -N2-methyl thiourea was recovered in the same manner as in 1-1 (1.9 g, 87%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.06 (d, 1H, J = 5.1 Hz), 6.81 (d, 1H, J = 5.1 Hz), 6.40 (br s, 1H), 6.00 (br s, 1H) , 3.74 (m, 2H), 3.05 (m, 2H), 2.89 (m, 3H), 2.20 (s, 3H).

(45-5) Synthesis of N1-2- (3-methylthiophen-2-ylethyl) -N2-methyl biguanide hydrochloride

The same method as in Step 1-2, except that N1-2- (3-methylthiophen-2-ylethyl) -N2-methyl thiourea (1.1 g, 5.10 mmol) prepared in step 45-4 was used. Recovery was carried out to recover N1-2- (3-methylthiophen-2-ylethyl) -N2-methyl biguanide hydrochloride (0.23 g, 16%).

¹ H NMR (300 MHz, D ₂ O) δ 7.02 (d, 1H, J = 5.1 Hz), 6.64 (d, 1H, J = 5.1 Hz), 3.04-3.06 (m, 2H), 2.72-2.74 (m, 2H), 2.48 (s, 3H), 1.93 (s, 3H).

Example 46 was carried out by the same method as Example 1, except for using the compound corresponding to the target compound in place of 3-methyl-2-thiophene-carbosaaldehyde in Step (45-1) of Example 45. Was prepared.

Example 3: N1-2- (thiophen-2-ylethyl) -N2-1-naphthyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.71-7.72 (m, 1H), 7.28-7.50 (m, 5H), 6.79-6.90 (m, 4H), 3.3.37-3.33 (m, 2H), 2.97-3.03 (t, 2H, J = 6.9)

Example 4: N1-2- (thiophen-2-ylethyl) -N2-hexyl biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.25-7.35 (m, 1H), 6.81-6.88 (m, 2H), 3.20-3.24 (m. 2H), 2.88-3.05 (m, 4H), 2.89- 2.99 (m, 4H), 1.11-1.32 (m, 4H), 0.75-0.78 (m, 3H)

Example 5: N1-2- (thiophen-2-ylethyl) -N2-methyl biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.31-7.32 (m, 1H), 6.87-6.95 (m, 2H), 3.27-3.32 (t, 2H, J = 7.2), 2.93-2.98 (t, 2H , J = 7.2), 2.63 (s, 3H); Mass (ESI) m / z 226.7 (M ⁺ )

Example 6: N1-2- (thiophen-2-ylethyl) -N2-cyclohexyl biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.36-7.39 (m, 1H), 6.90-7.00 (m, 2H), 3.25-3.24 (m, 2H), 2.97-3.05 (m, 2H), 2.23- 2.28 (m, 1 H), 1.66-1.90 (m, 5 H), 1.14-1.18 (m, 5H); Mass (ESI) m / z 294.7 (M ⁺ )

Example 7: N1-2- (thiophen-2-ylethyl) -N2-t-butyl biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.38-7.42 (m, 1H), 6.94-7.01 (m, 2H), 3.25-3.24 (m, 2H), 2.99-3.04 (m, 2H), 1.25- 1.30 (m, 9 H); Mass (ESI) m / z 268.7 (M ⁺ )

Example 8: N1-2- (thiophen-2-ylethyl) -N2-benzyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.24-7.48 (m, 6H), 6.90-7.01 (m, 2H), 4.34-4.35 (m, 2H), 2.98-3.22 (m, 4H); Mass (ESI) m / z 302.7 (M ⁺ )

Example 9: N1-2- (thiophen-2-ylethyl) -N2-2-ethylphenyl biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.23-7.38 (m, 6H), 6.90-6.91 (m, 2H), 3.27-3.39 (m, 4H), 2.98-3.00 (m, 2H), 2.77- 2.79 (m, 2 H); Mass (ESI) m / z 316.5 (M ⁺ )

Example 10: N1-2- (thiophen-2-ylethyl) -N2-methyl biguanide hydrochloride (isomer)

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.34-7.36 (m, 1H), 6.92-6.95 (m, 2H), 3.55-3.59 (t, 2H, J = 7.4), 3.27-3.28 (m, 3H ), 3.06-3.11 (t, 2H, J = 7.4); mp 185.0-199.5 ℃

Example 11: N1-2- (thiophen-2-ylethyl) -N2-methyl biguanide hydrochloride (isomer)

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.32-7.35 (m, 1H), 6.90-6.93 (m, 2H), 3.28-3.53 (m, 2H), 2.95-3.16 (m, 2H), 2.80 ( s, 3H),

Example 12 N1-2- (thiophen-2-ylethyl) -N2-methyl biguanide hydrochloride (isomer)

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.32-7.33 (m, 1H), 6.88-6.91 (m, 2H), 3.47-3.51 (m, 2H), 3.01-3.05 (m, 2H), 2.90 ( s, 3H).

Example 13: N1-2- (thiophen-2-ylethyl) -N2-propyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.30-7.40 (m, 1H), 6.89-6.83 (m, 2H), 3.34-3.45 (m, 2H), 2.93-3.01 (m, 4H), 1.41 -1.50 (m, 2H), 0.78-0.85 (m, 3H)

Example 14 N1-2- (thiophen-2-ylethyl) -N2- (3-trifluoromethyl) phenyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.47-7.22 (m, 5H), 6.92-6.86 (m, 2H), 3.38-3.40 (m, 2H), 2.97-3.02 (m, 2H); mp 129-131 ℃

Example 15 N1-2- (thiophen-2-ylethyl) -N2- (4-methoxy) phenyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.26-7.28 (m, 1H), 7.02-7.05 (m, 2H), 6.85-6.92 (m, 4H), 3.70 (s, 3H), 3.32-3.37 ( m, 2H), 2.98-3.10 (m, 2H); mp 127-129 ℃

Example 16: N1-2- (thiophen-2-ylethyl) -N2- (2,5-dimethoxy) phenyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.26-7.29 (m, 1H), 6.83-6.94 (m, 3H), 6.67-6.70 (2H), 3.62 (s, 3H), 3.68 (s, 3H) , 3.33 (t, 2H, J = 7.0), 2.98 (t, 2H, J = 7.1); mp 179-181 ° C

Example 17: N1-2- (thiophen-2-ylethyl) -N2- (3,4,5-trimethoxy) phenyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.25-7.23 (m, 2H), 6.91-6.85 (m, 3H), 3.65 (s, 6H), 3.55 (s, 3H), 3.36 (t, 2H, J = 7.0 Hz), 2.97 (t, 2H, J = 7.3 Hz); mp 92-94 ℃

Example 18 N1-2- (thiophen-2-ylethyl) -N2- (4-fluorophenyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.29-7.31 (m, 1H, J = 4.5 Hz), 7.14-7.11 (m, 4H), 6.94-6.89 (m, 2H), 3.34 (t, 2H, J = 7.5 Hz), 3.00 (t, 2H, J = 7.3 Hz); mp 83-85 ℃

Example 19 N1-2- (thiophen-2-ylethyl) -N2- (4-methylphenyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.28-7.31 (m, 1H), 7.13-7.16 (m, 2H), 6.88-6.96 (m, 4H), 3.38 (t, 2H, J = 7.2 Hz) , 3.01 (t, 2H, J = 7.2 Hz), 2.21 (s, 3H); mp 97-99 ℃

Example 20: N1-2- (thiophen-2-ylethyl) -N2-phenyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.30-7.24 (m, 3H), 7.10-7.07 (m, 3H), 6.94-6.88 (m, 2H), 3.38 (t, 2H, J = 7.3 Hz ), 3.01 (t, 2H, J = 7.2 Hz)

Example 21 N1-2- (thiophen-2-ylethyl) -N2-cyclopentyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.27-7.26 (m, 1H), 6.92-6.90 (m, 1H), 6.85-6.83 (m, 1H), 3.75-3.79 (m, 1H), 3.28 ( t, 2H, J = 7.2 Hz), 2.94 (t, 2H, J = 7.2 Hz), 1.82-1.73 (m, 2H), 1.60-1.53 (m, 2H), 1.49-1.33 (m, 4H)

Example 22 N1-2- (thiophen-2-ylethyl) -N2-cycloheptyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.28-7.30 (m, 1H), 6.86-6.94 (m, 2H), 3.48-3.51 (m, 1H), 3.28-3.31 (m, 2H), 2.98- 3.05 (m, 2H), 1.54-1.60 (m, 2H), 1.13-1.45 (m, 10H)

Example 23 N1-2- (thiophen-2-ylethyl) -N2-pentyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.33 (m, 1H), 6.90-6.95 (m, 2H), 3.49-3.55 (m, 2H), 3.21-3.40 (m, 2H), 3.00-3.15 (m, 2H), 1.22-1.50 (m, 6H), 0.79-0.87 (m, 3H).

Example 24 N1-2- (thiophen-2-ylethyl) -N2-methoxyethyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.32-7.33 (m, 1H), 6.93-6.95 (m, 1H), 6.86-6.88 (m, 1H), 3.36-3.33 (m, 4H), 3.21 ( s, 5H), 2.97-3.02 (m, 2H)

Example 25 N1-2- (thiophen-2-ylethyl) -N2-butyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.30 (m, 1H), 6.93-6.88 (m, 2H), 3.50 (m, 2H), 2.79-3.10 (m, 4H), 1.16-1.45 (4H ), 0.82-0.86 (m, 3 H).

Example 26: N1-2- (thiophen-2-ylethyl) -N2-butyl biguanide hydrochloride (isomer)

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.19-7.30 (m 1H), 6.93-6.88 (m, 2H), 3.30-3.40 (m, 2H), 2.70-3.29 (m, 4H), 1.15- 1.60 (m, 4 H), 0.83-0.88 (m, 3 H).

Example 27 N1-2- (thiophen-2-ylethyl) -N2- (3-methylphenyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.29-7.31 (m 1H), 7.13-7.16 (m, 2H), 6.88-6.95 (m, 4H), 3.37-3.41 (m, 2H), 2.99-3.03 (m, 2H), 2.21 (s, 3H)

Example 28 N1-2- (thiophen-2-ylethyl) -N2- (4-t-butylphenyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.27-7.30 (m, 3H), 7.01-7.04 (d, 2H, J = 6.9), 6.87-6.94 (m, 2H), 3.36-3.40 (m, 2H) , 2.98-3.03 (m, 2H), 1.14 (s, 9H); mp 215.2-224.1 ° C

Example 29: N1-2- (thiophen-2-ylethyl) -N2- (4-hexylphenyl) biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.34-7.36 (m, 1H), 7.04-7.11 (m, 4H), 6.87-6.97 (m, 2H), 3.32-3.43 (m, 2H), 2.91- 3.04 (m, 2H), 2.47-2.50 (m, 2H) 1.49-1.51 (m, 2H), 1.25-1.35 (m, 6H), 0.81-0.85 (s, 3H); mp 155.8-160.8 ℃

Example 30 N1-2- (thiophen-2-ylethyl) -N2- (4-methoxyphenyl) biguanide hydrochloride (isomer)

¹ H NMR (300MHz, D ₂ O) δ 7.25-7.27 (m, 1H), 7.02-7.05 (m, 2H), 6.84-6.92 (m, 4H), 3.66 (s, 3H), 3.36-3.40 (m , 2H), 2.97-3.01 (m, 2H); mp 98.6-99.2 ℃

Example 31: N1-2- (thiophen-2-ylethyl) -N2- (2,5-dimethoxyphenyl) biguanide hydrochloride

¹ H NMR (300MHz, D ₂ O) δ 7.33-7.34 (m, 1H), 6.91-6.97 (m, 4H), 6.65-6.70 (m, 1H), 3.73 (s, 3H), 3.70 (s, 3H ), 3.33-3.39 (t, 2H, J = 6.9), 2.95-3.03 (t, 2H, J = 6.9)

Example 32: N1-2- (thiophen-2-ylethyl) -N2- (3,5-dimethoxyphenyl) biguanide hydrochloride

¹ H NMR (300MHz, D ₂ O) δ 7.28-7.30 (m, 1H), 6.88-6.95 (m, 2H), 6.21-6.29 (m, 3H), 3.66 (s, 6H), 3.37-3.42 (t , 2H, J = 6.9), 2.99-3.04 (t, 2H, J = 6.9); mp 162.8-167.1 ℃

Example 33: N1-2- (thiophen-2-ylethyl) -N2- (4-bromophenyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.39-7.45 (m, 2H), 7.24-7.27 (m, 1H), 7.02-7.05 (m, 2H), 6.84-6.95 (m, 2H), 3.35-3.42 (m, 2H), 2.96-3.05 (m, 2H); mp 126.8-133.7 ℃

Example 34 N1-2- (thiophen-2-ylethyl) -N2- (3-bromophenyl) biguanide hydrochloride

¹ H NMR (300MHz, D ₂ O) δ 7.20-7.33 (m, 4H), 7.08-7.11 (m, 1H), 6.85-6.95 (m, 2H), 3.36-3.41 (m, 2H), 2.95-3.05 (m, 2H); mp 107.1-111.6 ℃

Example 35: N1-2- (thiophen-2-ylethyl) -N2- (4-phenylphenyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.57-7.63 (m, 4H), 7.38-7.44 (m, 2H), 7.19-7.36 (m, 4H), 6.83-6.96 (m, 2H), 3.38-3.45 (m, 2H), 2.90-3.07 (m, 2H); mp 145.8-150.6 ℃

Example 36: N1-2- (thiophen-2-ylethyl) -N2- (4-phenoxyphenyl) biguanide hydrochloride

¹ H NMR (300MHz, DMSO-d ₆ ) δ 7.27-7.37 (m, 3H), 7.09-7.14 (m, 3H), 6.88-6.94 (m, 6H), 3.38 (t, 2H, J = 7.2 Hz) , 3.01 (t, 2H, J = 7.2 Hz); mp 87.1-96.0 ℃

Example 37: N1-2- (thiophen-2-ylethyl) -N2- (4-bromobenzyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.43-7.50 (m, 2H), 7.22-7.28 (m, 1H), 7.09-7.17 (m, 2H), 6.75-6.93 (m, 2H), 4.23 (s , 2H), 3.29-3.33 (m, 2H), 2.93-3.03 (m, 2H); mp 102.1-103.5 ℃

Example 38: N1-2- (thiophen-2-ylethyl) -N2- (4-methoxybenzyl) biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.24-7.26 (d, 1H, J = 6.7 Hz), 7.11-7.16 (d, 2H, J = 6.7 Hz), 6.80-6.91 (m, 4H), 4.18 ( s, 2H), 3.66 (s, 3H), 3.28-3.33 (t, 2H, J = 7.2 Hz), 2.92-3.02 (t, 2H, J = 7.2 Hz)

Example 39: N1-2- (thiophen-2-ylethyl) -N2- (3,5-dichlorophenyl) biguanide hydrochloride

¹ H NMR (300MHz, D ₂ O) δ 7.21-7.27 (m, 2H), 7.10 (s, 2H), 6.88-6.95 (m, 2H), 3.34-3.40 (m, 2H), 3.03-3.08 (m , 2H)

Example 40 N1-2- (thiophen-2-ylethyl) -N2- (4-methylbenzyl) biguanide hydrochloride

¹ H NMR (300MHz, D ₂ O) δ 7.24-7.27 (m, 1H), 7.079-7.09 (m, 3H), 6.81-6.91 (m, 3H), 4.34-4.37 (m, 2H), 3.28-3.33 (m, 2H), 2.96-3.04 (m, 2H), 2.25 (s, 3H)

Example 41: N1-2- (thiophen-2-ylethyl) -N2-isopropyl vaguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.34-7.36 (m, 1H), 6.90-6.99 (m, 2H), 3.70-3.75 (m, 1H), 3.35-3.40 (m, 2H), 2.95 -3.00 (m, 2H), 1.06-1.08 (m, 6H)

Example 42: N1-2- (thiophen-2-ylethyl) -N2-isobutyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.30-7.32 (m, 1H), 6.90-6.95 (m, 2H), 3.34-3.40 (m, 2H), 3.99-3.04 (m, 2H), 2.90- 2.98 (m, 2H), 1.73-1.77 (m, 1H), 0.81-0.99 (m, 6H)

Example 43 N1-2- (thiophen-2-ylethyl) -N2-cyclopropylmethyl biguanide hydrochloride

¹ H NMR (300 MHz, D ₂ O) δ 7.30-7.31 (m, 1H), 6.91-6.94 (m, 2H), 3.36-3.41 (m, 2H), 3.05-2.97 (m, 4H), 0.94- 0.97 (m, 1H), 0.46-0.43 (m, 2H), 0.19-0.18 (m, 2H)

Example 44: N1-2- (thiophen-2-ylethyl) -N2-allyl biguanide hydrochloride

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.28-7.30 (m, 1H), 6.90-6.93 (m, 2H), 5.73-5.80 (m, 1H), 5.10-5.20 (m, 2H), 3.77 -3.78 (m, 2H), 3.39-3.43 (m, 2H), 2.99-3.04 (m, 2H); mp 97-99 ℃

Example 46 Synthesis of N1-2- (5-ethylthiophen-2-ylethyl) -N2-methyl biguanide hydrochloride

¹ H NMR (300MHz, D ₂ O) δ 6.40-6.43 (m, 2H), 3.07-3.09 (m, 2H), 2.67-2.69 (m, 2H), 2.46-2.49 (m, 5H), 0.91-0.96 (m, 3 H).

Test Example

About the compound obtained in the said Example, it measured according to the method described in the following test example. The following test was performed three times each with four plates in groups of Examples 1 to 9, Examples 10 to 12, Examples 13 to 22, Examples 23 to 39, and Examples 40 to 46, and an average value was calculated. Each plate was compared using 2 mM metformin as a control. In this case, although tested with the same concentration of metformin, there is a slight difference in the mean value for the test results depending on the respective plate measurement environment.

In Table 1 below, the cholesterol synthesis inhibitory ability, the ability to inhibit the synthesis of triglycerides and the glucose formation inhibitory ability may be lowered by toxicity when there is cytotoxicity, and the value is not recognized, and the value of the control group is a reference. To become a number. In the case of glucose intake capacity, it is measured by using insulin and control at the same time. If it is more than 200% in consideration of the deviation based on cell-based experiment, it is judged that there is glucose intake capacity.

Test Example 1: Measurement of cholesterol synthesis inhibitory ability

In order to measure cholesterol synthesis inhibitory ability, which is an important function of AMP-activated protein kinase (AMPK), compounds of the present invention were subjected to the following test using HepG2 cells (ATCC: American Type Culture Collection), a hepatocyte model.

First, HepG2 cells, a hepatocyte model, were cultured in a medium containing 1% serum (DMEM (Gibco), FBS (Gibco)) for 24 hours, and then the compounds of each Example were treated at 100 μm or 500 μm concentrations for 24 hours. The cells were then disrupted with crushed solution (0.1 M potassium phosphate, pH 7.4, 0.05 M NaCl, 5 mM cholic acid, 0.1% Triton X-100 (Sigma). The reaction solution (2 U / ml cholesterol oxidant, 2 U / ml peroxidase, 0.2 U / ml cholesterol esterase, 300 μM Amplex red (Molecular probe), a fluorescent factor, was added thereto, and reacted at 37 ° C. for 30 minutes. The amount of cholesterol formed was quantified by measuring at 560/590 nm (ex / em) wavelength with a fluorimeter.

Lower levels indicate the ability to inhibit cholesterol synthesis. In the case of the control group, the compound is judged to be superior to the control concentration when the compound is less than the value of the control group compared to the synthetic inhibitory ability shown in each plate at 2 mM concentration. For example, Example 1 shows a lower cholesterol synthesis amount than the control at a concentration of 100 uM can be determined that the effect is at least 20 times better.

Test Example 2: Determination of Triglyceride

HepG2 cells (ATCC: American Type Culture Collection) were incubated for 24 hours in a medium containing 1% serum (DMEM (Gibco), FBS (Gibco)), and then the compounds of each example were prepared at a concentration of 100 μm or 500 μm for 24 hours. After treatment the cells were disrupted with crushed solution (0.1 M potassium phosphate, pH 7.4, 0.05 M NaCl, 5 mM choline acid, 0.1% Triton X-100). The same volume of reaction solution (0.76 U / ml glycerol kinase (Asan), 151333 U / ml peroxidase (Asan), 22.2 U / ml glycerol oxidant (Asan), 300 μM Amplex red (Molecular Probe) was added and reacted for 30 minutes at 37 ° C. After the reaction, the amount of neutral fatty acid formed was measured at 560/590 nm (ex / em) wavelength using a fluorimeter.

Lower values indicate the ability to inhibit triglyceride synthesis. The control group is judged to be superior to the control concentration when the compound is less than the control group compared to the neutral fatty acid synthesis inhibitory ability shown in each plate at 2 mM concentration. For example, in the case of Example 1, since the use concentration is 100 μM, the neutral fatty acid synthesis amount is lower than that of the control group, and thus it may be determined that the effect is at least 20 times better.

Test Example 3: Measurement of glucose formation inhibitory ability

HepG2 cells (ATCC: American Type Culture Collection) were incubated in 10% serum hyperglycemic medium, and then transferred to serum-free hypoglycemic medium (DMEM (Gibco), FBS (Gibco)) to transfer the compounds of each example at 100 μm concentration for 24 hours. After treatment, the cells were incubated for 4 hours after treatment with 0.5 μCi ¹⁴ C-lactate (Amersham Pharmacia) and 10 mM L-lactate (Sigma). After incubation, the medium was removed and the cells washed with PBS. 0.1N NaOH was added to the washed cells and left at room temperature for 1 hour. After neutralizing with 1N HCl, the amount of glucose formed in the cells was measured by a liquid scintillation counter.

Lower levels indicate the ability to inhibit glucose formation. In the case of the control group, the compound was determined to be superior to the control concentration when the compound was below the value of the control group compared with the glucose formation inhibitory ability of each plate at the concentration of 2 mM. For example, in the case of Example 1, the use concentration is 100uM, showing a lower amount of glucose than the control group can be determined that the effect is at least 20 times better.

Test Example 4 Measurement of Glucose Absorption Capacity

The muscle cell model C2C12 (ATCC: American Type Culture Collection) was used to induce differentiation into muscle cells in 2% calf serum-containing medium (DMEM (Gibco), FBS (Gibco)) for 6 days. C2C12 cells differentiated into muscle cells were transferred to serum-free hypoglycemic medium (DMEM (Gibco), FBS (Gibco)) and the compounds of each example were treated with 100 μm concentration, followed by incubation for 24 hours by treatment with 1 μM of insulin (Sigma). It was. After incubation, 1 μCi ³ H-deoxy-glucose (Amersham Pharmacia) and 10 μM deoxy-glucose (Sigma) were treated at 37 ° C. for 15 minutes. After treatment, the medium was removed and the cells were washed twice with PBS (phosphate buffered saline). Washed cells were treated with 0.1N NaOH and neutralized with 1N HCl. The amount of glucose absorbed into the cells was measured with a liquid scintillation counter.

The higher the value, the stronger the ability to reduce insulin resistance. For example, in Example 1, since it showed a similar effect to the control at 100uM, it is determined that the effect is about 20 times better than the control in terms of concentration.

Test Example 5: Cytotoxicity Test

To measure cytotoxicity, LDH (Lactate dehydrogenase) method was used. That is, in order to use the passaged cells for experiments, cells were separated from the implant using trypsin-EDTA solution, and the plate (96 well microplate, manufactured by Falcon) was dispensed so that the number of cells per well was 1 × 10 ^4. It was. The aliquoted cells were incubated in a CO ₂ incubator for 24 hours and attached to the bottom, and then the culture solution was removed with an aspirator, and the compound solution diluted in the culture solution with the compound and the control drug was used in the well containing the cells. 100 uL each was added in triplicate, and further incubated for 24 hours. Dimethyl sulfoxide (DMSO) was used as needed to dissolve the compound. After 24 hours of treatment, the supernatant medium and LDH (Roche) reagent were mixed and reacted, and then measured at the reaction absorbance (490 nm) to determine cytotoxicity.

In the case of cytotoxicity, it is generally determined that there is cytotoxicity at 30% or more at the concentration. Therefore, embodiments showing a value of 30% or less can be safely used as a drug because there is no cytotoxicity.

The results are shown in Table 1 below.

Claims (12)

N1-2-thiophen-2-ylethyl-N2-substituted biguanide derivative compound of Formula 1, or a pharmaceutically acceptable salt thereof. [Formula 1]

Where R is C ₁ -C ₈ alkyl, allyl, C ₁ -C ₈ alkoxyalkyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkylC ₁ -C ₃ alkyl, optionally substituted phenyl or phenylC _1- C ₃ alkyl, optionally substituted naphthyl or naphthylC ₁ -C ₃ alkyl, or adamantyl group; R ₁ is hydrogen or a C ₁ -C ₆ alkyl group. The method of claim 1, In Formula 1, R is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, allyl, Methoxymethyl, methoxyethyl, methoxyethoxyethoxyethyl, ethoxymethyl and octyloxymethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopropanemethyl, cyclobutanemethyl, cyclopentanemethyl Cyclohexanemethyl, cyclopropaneethyl, cyclobutaneethyl, cyclopentaneethyl, cyclohexaneethyl, cyclopropanepropyl, cyclobutanepropyl, cyclopentanepropyl, cyclohexanepropyl, and unsubstituted or substituted benzyl, phenyl, Naphthyl, 1-phenylethyl, 2-phenylethyl, 1-phenylpropyl, 2-phenylpropyl, 3-phenylpropyl, 1-naphthylmethyl, 2-naphthylmethyl, 1-naphthylethyl and 2-naphthyl Selected from the group consisting of ethyl , In this case, the substituents may be each the same or different from one to six, halogen atoms, a hydroxy group, a nitro group, a cyano group, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl group, C ₁ -C ₆ cycloalkyl Alkyl group, C ₆ -C ₁₀ aryl group, C ₆ -C ₁₀ aryloxy group, C ₁ -C ₆ alkoxy group, C ₁ -C ₆ haloalkoxy group, C ₃ -C ₆ cycloalkyloxy group, C ₁ -C ₇ alkanoyl group, carboxyl group, carbamoyl group, alkylamino group, C ₂ -C ₇ sulfonic acid group, sulfonamido group and C ₁ -C ₆ alkylthio group; A compound of formula (1) or a pharmaceutically acceptable salt thereof, wherein R ₁ is hydrogen, methyl, ethyl, propyl, butyl, pentyl or hexyl. The method of claim 2, R is ethyl, t-octyl, 1-naphthyl, hexyl, methyl, cyclohexyl, t-butyl, benzyl, 2-ethylphenyl, propyl, 3-trifluoromethylphenyl, 4-methoxyphenyl, 2,5 Dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 4-fluorophenyl, 4-methylphenyl, phenyl, cyclopentyl, cycloheptyl, pentyl, methoxyethyl, butyl, 3-methylphenyl, 4-t -Butylphenyl, 3-hexylphenyl, 4-methoxyphenyl, 2,5-dimethoxyphenyl, 3,5-dimethoxyphenyl, 4-bromophenyl, 3-bromophenyl, 3-phenylphenyl, 3-phenoxyphenyl, 4-bromobenzyl, 4-methoxybenzyl, 3,5-dichlorophenyl, 4-methylbenzyl, isopropyl, isobutyl, cyclopropylmethyl, 2- (2- (2-meth Methoxyethoxy) ethoxy) ethyl or allyl; A compound of formula 1, or a pharmaceutically acceptable salt thereof, wherein R ₁ is hydrogen, 3-methyl or 5-ethyl. The method of claim 1, The pharmaceutically acceptable salts are formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, Acids selected from the group consisting of glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid It is a salt of the compound of formula (1) or a pharmaceutically acceptable salt thereof. 1) reacting a compound of Formula 2 with NaBH ₄ to obtain a compound of Formula 3; 2) reacting a compound of Formula 3 with thionyl chloride in an organic solvent to obtain a compound of Formula 4; 3) reacting a compound of Formula 4 with sodium cyanide in dimethyl sulfoxide (DMSO) to obtain a compound of Formula 5; 4) reacting the compound of Formula 5 with sodium borohydride and nickel chloride hydrate to obtain a compound of Formula 6; 5) reacting a compound of Formula 6 with R-isothiocyanate (RNCS) in an organic solvent in the presence of a base to obtain a compound of Formula 8; And 6) A process for preparing a compound of formula 1, comprising reacting a compound of formula 8 in a guanidine solution in the presence of mercury oxide: [Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Chemical Formula 5]

[Formula 6]

[Formula 8]

In the above formulas, R and R ₁ are as defined in claim 1. 1) reacting a compound of Formula 2 with NaBH ₄ to obtain a compound of Formula 3; 2) reacting a compound of Formula 3 with thionyl chloride in an organic solvent to obtain a compound of Formula 4; 3) reacting a compound of Formula 4 with sodium cyanide in dimethyl sulfoxide (DMSO) to obtain a compound of Formula 5; 4) reacting the compound of Formula 5 with sodium borohydride and nickel chloride hydrate to obtain a compound of Formula 6; 5) reacting a compound of formula 6 with ethylchloroformate in an organic solvent in the presence of carbon disulfide and a base to obtain a compound of formula 7; 6) reacting a compound of Formula 7 with NH ₂ R in the presence of a base to obtain a compound of Formula 8; And 7) A process for preparing a compound of formula 1 comprising reacting a compound of formula 8 in a guanidine solution in the presence of mercury oxide: [Formula 1]

[Formula 2]

[Formula 3]

[Formula 4]

[Chemical Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

In the above formulas, R and R ₁ are as defined in claim 1. The method according to claim 5 or 6, The base is selected from the group consisting of triethylamine, trimethylamine and diisopropylethylamine, and the organic solvent is selected from the group consisting of dichloromethane, dichloroethane and dimethylformamide. A pharmaceutical composition for preventing or treating diabetes, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for preventing or treating metabolic syndrome, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The method of claim 9, Pharmaceutical composition, characterized in that the metabolic syndrome is insulin-independent diabetes, obesity or atherosclerosis. A pharmaceutical composition for the prophylaxis or treatment of a cancer lacking the gene P53, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The method according to any one of claims 8 to 11, The pharmaceutically acceptable salts are formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, Acids selected from the group consisting of glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid Pharmaceutical composition, characterized in that the salt.